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Fig. 2 | BMC Biology

Fig. 2

From: Functional role of dimerization and CP190 interacting domains of CTCF protein in Drosophila melanogaster

Fig. 2

a Schematic drawing of luciferase reporter constructs. b Firefly luciferase expression from the five reporters shown in a when co-transfected with empty vector, with a vector encoding a 3xFLAG-tagged-(nuclear localization signal)-LexA fusion protein, or with a vector encoding the 3xFLAG-tagged N-terminal dCTCF-(nuclear localization signal)-LexA fusion protein. A plasmid encoding the Renilla luciferase under the control of the actin promoter was used to correct for variations in transfection efficiency, and expression of the firefly luciferase was normalized in each case to Renilla luciferase. Each transfection experiment was performed in three independent biological replicates and each lysate was measured in four technical replicates. Error bars show standard deviations of measurements of all summarized replicates. c Chromatin immunoprecipitation of S2 cells co-transfected with the 4xLesA TATA-box reporter or the basic promoterless reporter and either of two fusion protein expression constructs, the 3xFLAG-tagged (nuclear localization signal) LexA construct or the 3xFLAG-tagged-N-terminal dCTCF-(nuclear localization signal)-LexA construct. Fixed and processed S2 chromatin samples were immunoprecipitated with antibodies directed against (as indicated) the dCTCF N-terminus, the dCTCF C-terminus, CP190, or FLAG, and then assayed for the presence of sequences corresponding to the 4xLexA TATA reporter or the basic reporter constructs as indicated. Each chromatin immunoprecipitation experiment was performed in three independent biological replicates. Error bars show standard deviations of summarized biological replicates after quadruplicate PCR measurements in each experiment. The results are presented as a percentage of input DNA. Basic no promoter, bla basic promoterless reporter, Hsp70 firefly luciferase with an hsp70 promoter, TATA firefly luciferase with a minimal TATA-box promoter, 4xlex bs, firefly luciferase with four copies of the LexA recognition sequence, 4xlex bs + TATA, firefly luciferase with four copies of the LexA recognition sequence linked to a minimal TATA-box promoter

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